Team:UT Dallas/week7

From 2013.igem.org

8/18

<a name="h.ifbebkzetykw"></a>Miniprep/glycerol stock

<a name="h.mnmnvk188va8"></a>i5-38-9 → i5-40-1

<a name="h.scsrhbifihao"></a>i5-38-10 → i5-40-2

<a name="h.i2po0dnsdx3n"></a>ComA → 114.6

<a name="h.m2wdryonbfiu"></a>

<a name="h.to30bq1m0q5u"></a>Transformation

<a name="h.l3409m1ri7qd"></a>i5-40-3 3:1 → 0 colonies

<a name="h.ocidj62r5isw"></a>i5-40-4 0:1 → 0 colonies

<a name="h.vb1li239hfaw"></a>+ control → 0 colonies

<a name="h.dm2t73uh0qw6"></a>

<a name="h.o73u54952308"></a>8/19

<a name="h.pn9yzbf71qtt"></a>Redo of Transformation from 8/16

<a name="h.flbywvs88xxr"></a>1→ i5-40-5 (3:1 CP medium -Cra-pFru-RBS-GFP) → 5 colonies → i5-40-10 → ruined (left out)

<a name="h.nsybqxkz5g7a"></a>                                                                i5-40-11

<a name="h.4juaxuwaonps"></a>2 → i5-40-6 (0:1) → 10 colonies

<a name="h.iuhk8upurx6h"></a>3→ i5-40-7 (3:1 CP-Scr-pScr-RBS-GFP) → 3 colonies → i5-40-12

<a name="h.36suk8lxoph3"></a>                                                        i5-40-13

<a name="h.6fj4n9fmu7z1"></a>4→ i5-40-8 (0:1) → 2 colonies

<a name="h.lxrvrcmnexo"></a>i5-11-5 → positive control → 150+ colonies

<a name="h.mgghxdgne91n"></a>

<a name="h.ufar1jn6p8at"></a>Digestion

<a name="h.d5fk1zvfvmmv"></a>

<a href="#" name="7fc8a7ed55d7e1111ec76c50c6270bf248e7d445"></a><a href="#" name="63"></a><tbody></tbody>

<a name="h.sa6ktn6duges"></a>

<a name="h.bp9lsvyr2m66"></a>i5-11-5

<a name="h.33qe6za2soee"></a>DNA

<a name="h.p799cfyt5w5n"></a>30

<a name="h.w462eorzimo1"></a>Buffer

<a name="h.dsuw5q3ktux7"></a>5

<a name="h.pm1x329bppvf"></a>XbaI

<a name="h.35bvbcrh04at"></a>1

<a name="h.dqq70wnorwst"></a>SpeI

<a name="h.n8t060hzb0rv"></a>1

<a name="h.yfqzs5j2yygr"></a>Water

<a name="h.cd0vz7ker88p"></a>13

<a name="h.y34ag28u915w"></a>Total

<a name="h.qa7a6h6vqjt1"></a>50

<a name="h.n7lx4h61eqsl"></a>→ i5-40-9 → 9.3 ng/ul

<a name="h.5ikwccmkxs9d"></a>

<a name="h.s4y8gz6dabcw"></a>Test Digestion

<a name="h.tyka6safzh5u"></a> 

<a href="#" name="450cc4705bd2eb4263286660d3172c77ec4e8d94"></a><a href="#" name="64"></a><tbody></tbody>

<a name="h.l5zrvrv86pgc"></a>

<a name="h.wdsxvj91amrx"></a>i5-40-1

<a name="h.jd1vra7uhskr"></a>i5-40-2

<a name="h.8sllm73qqti7"></a>DNA

<a name="h.puwts5wbv6lb"></a>2

<a name="h.v5ge72wgqvrr"></a>3

<a name="h.ua2tqjhwrqxk"></a>Buffer

<a name="h.7fy7azaxjcls"></a>2

<a name="h.mhlr49jsqele"></a>2

<a name="h.rmwv4pxazb31"></a>EcoRI

<a name="h.dk64vkk8s8q8"></a>1

<a name="h.xp6vhzcviv2"></a>1

<a name="h.vephoqgtac8y"></a>SpeI

<a name="h.oveohsy90059"></a>1

<a name="h.q7k3su7m8sce"></a>1

<a name="h.vmjlzbasdzfw"></a>Water

<a name="h.bl3va9a8zi85"></a>14

<a name="h.fht3tz979i7c"></a>13

<a name="h.7u3ay1sgg7h7"></a>Total

<a name="h.8hgnysqhmca0"></a>20

<a name="h.m1yye8xsazlh"></a>20 (trashed)

<a name="h.1z2w74lqafob"></a>

<a name="h.sl2xp8r0vpvf"></a>8/20

<a name="h.4m0iignjnoym"></a>Redo of digestion from 8/16

<a name="h.kxkvhlsg82hq"></a>

<a href="#" name="576aa37909ccbc7ab78bb39d396a04524955882b"></a><a href="#" name="65"></a><tbody></tbody>

<a name="h.am08x4lceuvm"></a>

<a name="h.8bxxao1a9oz1"></a>i5-35-5

<a name="h.z5yl6t6x0iya"></a>i5-35-3

<a name="h.o2709bj1tqz4"></a>DNA

<a name="h.glmbqo5yz0bi"></a>20

<a name="h.dz0hpw8nws75"></a>20

<a name="h.g2pzidzem40x"></a>NEBuffer

<a name="h.1qklhzz665gb"></a>5

<a name="h.1uhprsiyop7z"></a>5

<a name="h.89eqqansw97o"></a>PstI

<a name="h.q74l5h5ron8p"></a>1

<a name="h.o378wbivoc3z"></a>1

<a name="h.u38ue4h7stxj"></a>SpeI

<a name="h.fxb4w9a5uyly"></a>1

<a name="h.cixchzlpt22q"></a>1

<a name="h.3flm06yf9h8i"></a>Water

<a name="h.31f0bbd4iagv"></a>13

<a name="h.ka02dlala5a"></a>13

<a name="h.1n9hgf8lsgdf"></a>Total

<a name="h.3e5hyuvdiqlv"></a>50

<a name="h.qlu0l1vsnuzg"></a>50

<a name="h.5xue9dxt1fnm"></a>i5-35-5 → i5-41-1 (BB-CP-Cra) → 18.9 ng/ul

<a name="h.2gb255mrui5z"></a>i5-35-3 → i5-41-2 (BB-CP-Scr) → 62.5

<a name="h.67xki8v9adnl"></a>

<a name="h.hr1wpj2sashf"></a>Ligation

<a name="h.chk8ysgm69qh"></a>Vector: i5-40-9

<a name="h.o0r3kivu93yo"></a>Insert: i5-36-2

<a name="h.jo8twjy763jx"></a>

<a href="#" name="f50d59901cd2c5fe1f4537018173a0c0cea8fb74"></a><a href="#" name="66"></a><tbody></tbody>

<a name="h.5pd7k6gx4gtv"></a>

<a name="h.ycpvcibi7m62"></a>3:1

<a name="h.7mhyiwb57p4s"></a>0:1

<a name="h.w3zryvkyovst"></a>Vector

<a name="h.w0fjvtdkikgf"></a>2.5

<a name="h.3z4hsmmufgtj"></a>2.5

<a name="h.1vbwubt4kq64"></a>insert

<a name="h.53st91cnj16h"></a>11.5

<a name="h.uhgrnbhrobgw"></a>0

<a name="h.w7y1tof7tcy5"></a>Buffer

<a name="h.pfbn3egcritq"></a>3

<a name="h.do2n43gs0jj3"></a>3

<a name="h.4xhrnsr62yek"></a>Ligase

<a name="h.ylyr9flqjnz3"></a>1

<a name="h.bs7vr5jn6vrw"></a>1

<a name="h.93d3ohc6ene9"></a>Water

<a name="h.z7bumvk926a3"></a>3

<a name="h.644me238kqk2"></a>14.5

<a name="h.oak2arqxxvl8"></a>Total

<a name="h.dbvmeni66pr1"></a>20

<a name="h.blii8asauyun"></a>20

<a name="h.nb4lxb1slou"></a>

<a name="h.kf9qi4q3nga4"></a>1

<a name="h.q3783zb39yp2"></a>2

<a name="h.g3l0yp6unyzj"></a>

<a name="h.tmboy0viwsd8"></a>Mutagenesis

<a name="h.rnzf91b5sw64"></a>i5-40-1

<a name="h.bsyyuavb4x2a"></a>

<a href="#" name="dd55b66ddda0522c755edc6a490d67bc0618db7a"></a><a href="#" name="67"></a><tbody></tbody>

<a name="h.o1rk9loosjpy"></a>QS

<a name="h.ttgcky750ysg"></a>25

<a name="h.mjzu4gnove5q"></a>F primer

<a name="h.5b54kguvhgek"></a>1.1

<a name="h.p02py95xlaw0"></a>R primer

<a name="h.nbkchwasx1re"></a>1.1

<a name="h.usdykjjv16yj"></a>DNA

<a name="h.3fk3ftv5mbfn"></a>1

<a name="h.1pe8vwlfqj0n"></a>Water

<a name="h.fe07za1ikr66"></a>21.8

<a name="h.996udgk7d9tv"></a>Total

<a name="h.hj8bf0n2py0x"></a>50

<a name="h.4yy3qw29fran"></a>

<a name="h.hy496lyu2byi"></a>1

<a name="h.37hdz8y85vt6"></a>

<a name="h.2jo8equj3y4j"></a>Transformation

<a name="h.b8d4boty1ozi"></a>i5-42-2 3:1 → 0 colonies

<a name="h.jf48uef5u2cc"></a>i5-42-3 0:1 → 0 colonies

<a name="h.ug09b9giy3ce"></a>i5-11-5 + control → 50+ colonies

<a name="h.ff1h7nstiw56"></a>

<a name="h.vly2gyflodyy"></a>Ligation

<a href="#" name="5a5eca30e97f14e6ecc746d2e59e0cfbaab43c92"></a><a href="#" name="68"></a><tbody></tbody>

<a name="h.erte45s39r53"></a>

<a name="h.cbzh5ihftoaf"></a>Insert: i5-38-7 (pFru-RBS-GFP)

<a name="h.5dkloqhqe0pl"></a>Vector: i5-41-1 (BB-CP-Cra)

<a name="h.9i3e8je9olnv"></a>

<a name="h.z9yds5oaayup"></a>3:1

<a name="h.eg7loxgt1gcz"></a>0:1

<a name="h.3ri8oh5zwpru"></a>Insert

<a name="h.gyptirvkhvka"></a>4.4

<a name="h.fjrggskzwase"></a>0

<a name="h.og3yfff8jn8z"></a>Vector

<a name="h.lg989dp37cix"></a>2.7

<a name="h.g1mzx4lckuxa"></a>2.7

<a name="h.gwd6kvhdwtll"></a>Buffer

<a name="h.7t7j3ftuze84"></a>2

<a name="h.bt9hasw3x1st"></a>2

<a name="h.4xnctsup183o"></a>Ligase

<a name="h.i16hd93qnth7"></a>1

<a name="h.5e20a1s2ryhw"></a>1

<a name="h.uyd45ib8bkyo"></a>Water

<a name="h.v2z6iuusnzhf"></a>9.9

<a name="h.ujnrtuf2d4xm"></a>14.3

<a name="h.k971rkn8ail7"></a>Total

<a name="h.36yan3xkgmx8"></a>20

<a name="h.gheriwbuuxlm"></a>20

<a name="h.5pbolx6vnfh9"></a>

<a name="h.i0ylwjsusvo5"></a>1

<a name="h.w1nhxtxg5u0"></a>2

<a name="h.yai690lycdne"></a>

<a href="#" name="941e2e97bad5d7e73b995a6a4d53fde57e9bc6b1"></a><a href="#" name="69"></a><tbody></tbody>

<a name="h.6cc44yqku5ih"></a>

<a name="h.u506dn9xub6x"></a>insert: i5-38-8 (pScr-RBS-GFP)

<a name="h.drslfp7u5rcm"></a>Vector: i5-41-2 (BB-CP-Scr)

<a name="h.27nphgsg5pl6"></a>

<a name="h.5aqtu7wslugl"></a>3:1

<a name="h.3mnese3kbkwj"></a>0:1

<a name="h.e3cjfs688p7v"></a>insert

<a name="h.emvob8f710uk"></a>9.2

<a name="h.txz8b9mpwwxv"></a>0

<a name="h.gw2yjifvkppr"></a>Vector

<a name="h.k95k6am3j6w2"></a>8

<a name="h.wqwby3kliwbs"></a>8

<a name="h.2l6pjjnexi8q"></a>Buffer

<a name="h.j0bdrrjwv7rh"></a>2

<a name="h.i4g01gs957c8"></a>2

<a name="h.lk8ya4esnxs8"></a>Ligase

<a name="h.14jq802rt6r1"></a>1

<a name="h.p8nyqdn2k99f"></a>1

<a name="h.17scjvz8ulij"></a>Water

<a name="h.7gm326h2pemd"></a>7

<a name="h.8jb2u84o2zm8"></a>18.2

<a name="h.x1ekoloiefuu"></a>Total

<a name="h.58wk5pl14q8o"></a>20

<a name="h.48153rl7rvb"></a>20

<a name="h.919jxfr62q9y"></a>

<a name="h.8fgjexlp2ui8"></a>3

<a name="h.a7yxfza15mmh"></a>4

<a name="h.n2lx76v5va1j"></a>

<a name="h.8lynof1u9x03"></a>Transformation

<a name="h.snw66067sgnd"></a>1→ i5-42-4 (3:1 CP-Cra-pFru-RBS-GFP) → 20 colonies → picked 2 colonies → i5-42-10

<a name="h.afb7pxmhnvj5"></a>                                                                                i5-42-11

<a name="h.b57uw6l2sjgu"></a>2→ i5-42-5 (0:1) → 2 colonies

<a name="h.qq9o6jebmfll"></a>3→ i5-42-6 (3:1 CP-Sur-pScr-RBS-GFP) → 0 colonies

<a name="h.9cvseccq8tqy"></a>4→ i5-42-7 (0:1) → 0 colonies

<a name="h.lh376a5ks45g"></a>

<a name="h.udj8mnoub7qi"></a>i5-40-1 → i5-42-8 (Com A)

<a name="h.nupjgtj83emx"></a>i5-11-5 → i5-42-9 (+ control)

<a name="h.i4vso9ak72hj"></a>

<a name="h.2xzf2h6umj5k"></a>8/21

<a name="h.3w8i4ewalkni"></a>Redo transformation from 8/20

<a name="h.hnqyiixn1tsk"></a>i5-43-1 (3:1) →

<a name="h.7otbt4oecxsx"></a>i5-43-2 (0:1) →

<a name="h.fe2piguhq46b"></a>+ control i5-11-5 →

<a name="h.9x79t2pa2xhj"></a>3 → i5-43-4 (3:1 CP-Scr-pScr-RBS-GFP) → 1 colony → i5-43-12

<a name="h.gbgw6a67iom5"></a>4→ i5-43-5 (0:1)

<a name="h.7x77io36l3rk"></a>

<a name="h.unj1p1ov0t3w"></a>Miniprep and glycerol stock (10,12)

<a name="h.jh5k3ar45w6q"></a>i5-40-10 [CP-Fru-pFru-RBS-GFP] i5-40-10→ i5-43-6 → 60.7

<a name="h.160be9e4szah"></a>i5-40-11  [CP-Fru-pFru-RBS-GFP] i5-40-11 → i5-43-7→ 75.6

<a name="h.sg5xpvccmfve"></a>i5-40-12 [CP-Sru-pSru-RBS-GFP] i5-40-12 → i5-43-8→ 50.7

<a name="h.dbqq5vocz98e"></a>i5-40-13 [CP-Sru-pSru-RBS-GFP] i5-40-13→ i5-43-9→ 80.3

<a name="h.ibbcfzj04pp9"></a>

<a name="h.1pw3gy7kre22"></a>Test Digestion

<a name="h.jjyje9wczd7r"></a>

<a href="#" name="73b72c965d49da87cff5ef97b7099d6d0f26e612"></a><a href="#" name="70"></a><tbody></tbody>

<a name="h.y61vs04rmj23"></a>

<a name="h.8cb8w1ggiprf"></a>i5-43-6

<a name="h.9z13sctj0yp0"></a>i5-43-7

<a name="h.1gl2j6j1bos"></a>i5-43-8

<a name="h.96gi2ejgam4s"></a>i5-43-9

<a name="h.qx0cj9aiozfx"></a>DNA

<a name="h.xjgyl9m4y8lq"></a>4

<a name="h.gj1n1gjqss5"></a>3

<a name="h.7a35r7it0jlk"></a>4

<a name="h.m92slhxo5mz3"></a>3

<a name="h.gdyvtu4yzpfv"></a>Buffer

<a name="h.xofs3nh8i4jp"></a>2

<a name="h.lvkumyc7v44x"></a>2

<a name="h.dqjnyd6qzbv2"></a>2

<a name="h.jy9i4vro4rc0"></a>2

<a name="h.78esrv842nxh"></a>XbaI

<a name="h.icpp1x7t5b1h"></a>1

<a name="h.ix53yfuupabf"></a>1

<a name="h.etmvih43ov1k"></a>1

<a name="h.yrim4o6ycjv6"></a>1

<a name="h.b4vu07wff1w1"></a>PstI

<a name="h.3bmjq2k142r7"></a>1

<a name="h.icf05wcwcm3k"></a>1

<a name="h.skt7zoha9kl9"></a>1

<a name="h.n0g4snmrqgif"></a>1

<a name="h.c8hws67jxjs4"></a>Water

<a name="h.19t4gbmygs8v"></a>12

<a name="h.2le8re1z4oau"></a>13

<a name="h.i3km7wkx7tlj"></a>12

<a name="h.ytser0mhfrj7"></a>13

<a name="h.1nux9bodrn68"></a>Total

<a name="h.gvr0qfpfhx9"></a>20

<a name="h.ab63tpdb0hx3"></a>20

<a name="h.qwudz3ltiehu"></a>20

<a name="h.e2l4z6ec097u"></a>20

<a name="h.4stnwecbfya0"></a>

<a name="h.whnluhjrkk6z"></a>1

<a name="h.f402t6w44eoh"></a>2

<a name="h.6zp03dk8s385"></a>3

<a name="h.93otocnfz340"></a>4

<a name="h.s08d8mo06kj5"></a>

<a name="h.ud5cbxq0ovd"></a>

<a name="h.ayv1jfa0dy6q"></a>Redo Transformation

<a name="h.jopkxd4m76sd"></a>i5-42-8 → i5-43-10 (comA) → infinite colonies → picked 2 → i5-43-2, i5-43-3

<a name="h.ycpacih76ncj"></a>i5-11-5 → i5-43-11 (+ control)

<a name="h.5gadth3xdr6r"></a>

<a name="h.awlj0d13lrvz"></a>Redo Digestion from 8/20

<a name="h.dqf52egpqluz"></a>

<a name="h.jjyje9wczd7r"></a>

<a href="#" name="566c6151e072578eb7cac42ed9e2dc3af5f2b7c5"></a><a href="#" name="71"></a><tbody></tbody>

<a name="h.y61vs04rmj23"></a>

<a name="h.8cb8w1ggiprf"></a>i5-35-5

<a name="h.9z13sctj0yp0"></a>i5-35-3

<a name="h.qx0cj9aiozfx"></a>DNA

<a name="h.xjgyl9m4y8lq"></a>20

<a name="h.gj1n1gjqss5"></a>20

<a name="h.gdyvtu4yzpfv"></a>Buffer

<a name="h.xofs3nh8i4jp"></a>5

<a name="h.lvkumyc7v44x"></a>5

<a name="h.78esrv842nxh"></a>PstI

<a name="h.icpp1x7t5b1h"></a>2.5

<a name="h.ix53yfuupabf"></a>2.5

<a name="h.b4vu07wff1w1"></a>SpeI

<a name="h.3bmjq2k142r7"></a>2.5

<a name="h.icf05wcwcm3k"></a>2.5

<a name="h.c8hws67jxjs4"></a>Water

<a name="h.19t4gbmygs8v"></a>20

<a name="h.2le8re1z4oau"></a>20

<a name="h.1nux9bodrn68"></a>Total

<a name="h.gvr0qfpfhx9"></a>50

<a name="h.ab63tpdb0hx3"></a>50

<a name="h.4stnwecbfya0"></a>

<a name="h.whnluhjrkk6z"></a>1

<a name="h.f402t6w44eoh"></a>2

<a name="h.k24kbmhdovu7"></a>

<a name="h.2ejv7cq2k11t"></a>PCR

<a name="h.jjyje9wczd7r"></a>

<a href="#" name="6b8c206b6f865f4a5bdcc236629f4d7dadba74c0"></a><a href="#" name="72"></a><tbody></tbody>

<a name="h.8cb8w1ggiprf"></a>Q5

<a name="h.9z13sctj0yp0"></a>25

<a name="h.xjgyl9m4y8lq"></a>F

<a name="h.gj1n1gjqss5"></a>2.5

<a name="h.xofs3nh8i4jp"></a>R

<a name="h.lvkumyc7v44x"></a>2.5

<a name="h.icpp1x7t5b1h"></a>DNA (i5-12-1)

<a name="h.ix53yfuupabf"></a>1

<a name="h.3bmjq2k142r7"></a>Water

<a name="h.icf05wcwcm3k"></a>19

<a name="h.19t4gbmygs8v"></a>Total

<a name="h.2le8re1z4oau"></a>50

<a name="h.gvr0qfpfhx9"></a>

<a name="h.ab63tpdb0hx3"></a>1

<a name="h.7lvqgxsddpzc"></a>

<a name="h.btaulqmd424o"></a>Purification

<a name="h.igdjx1blp16b"></a>1 → 37.5

<a name="h.jldlygwz4342"></a>

<a name="h.k0552b3lp75r"></a>Digestion

<a name="h.jjyje9wczd7r"></a>

<a href="#" name="db938a0c00d6ed48a8ab642cd9bfcfc260b6d3c2"></a><a href="#" name="73"></a><tbody></tbody>

<a name="h.8cb8w1ggiprf"></a>DNA

<a name="h.9z13sctj0yp0"></a>30

<a name="h.xjgyl9m4y8lq"></a>Buffer

<a name="h.gj1n1gjqss5"></a>5

<a name="h.xofs3nh8i4jp"></a>XBaI

<a name="h.lvkumyc7v44x"></a>2.5

<a name="h.icpp1x7t5b1h"></a>SpeI

<a name="h.ix53yfuupabf"></a>2.5

<a name="h.3bmjq2k142r7"></a>Water

<a name="h.icf05wcwcm3k"></a>10

<a name="h.jjyje9wczd7r"></a>

<a href="#" name="dae6cd2e2372b959aaaba6f0a24d69becfca22a8"></a><a href="#" name="74"></a><tbody></tbody>

<a name="h.19t4gbmygs8v"></a>Total

<a name="h.2le8re1z4oau"></a>50

<a name="h.4ruwoq3b0pf"></a>→ i5-45-1

<a name="h.42aytlk03fit"></a>

<a name="h.iqmttgmi1z5"></a>8/22

<a name="h.8lkz2wl2gqjf"></a>Ligation

<a name="h.rx3cnfg6na3b"></a>Insert: i5-38-7

<a name="h.7y3v88mnqo5j"></a>Vector: i5-44-1